LOCUS       62056_12795        8794 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8794)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     intron          2100..3111
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     promoter        3321..3532
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3545..3813
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3869..4534
                     /codon_start=1
                     /label=CopGFP
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
                     /translation="LPAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             4604..4657
                     /codon_start=1
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4658..5254
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    5365..5497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6773..6789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6797..6813)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6821..6851)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6866..6887)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7175..7763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7937..8794)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc caaggcctac tagtattatg cccagtacat
     1381 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1441 ggacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     1501 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     1561 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     1621 actttccatt gacgtcaatg ggtggactat ttacggtaaa ctgcccactt ggcagtacat
     1681 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     1741 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     1801 ttagtcatcg ctattaccat gggtcgaggt gagccccacg ttctgcttca ctctccccat
     1861 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
     1921 gatgggggcg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
     1981 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
     2041 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
     2101 gagtcgctgc gttgccttcg ccccgtgccc cgctccgcgc cgcctcgcgc cgcccgcccc
     2161 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
     2221 gctgtaatta gcgcttggtt taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc
     2281 ttaaagggct ccgggagggc cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg
     2341 tgtgtgtgcg tggggagcgc cgcgtgcggc ccgcgctgcc cggcggctgt gagcgctgcg
     2401 ggcgcggcgc ggggctttgt gcgctccgcg tgtgcgcgag gggagcgcgg ccgggggcgg
     2461 tgccccgcgg tgcggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     2521 ggggggtgag cagggggtgt gggcgcggcg gtcgggctgt aacccccccc tgcacccccc
     2581 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtgcgg ggcgtggcgc
     2641 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
     2701 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc cccggagcgc cggcggctgt
     2761 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     2821 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     2881 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     2941 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     3001 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     3061 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     3121 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattctgcag tcgacggtac
     3181 ccgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     3241 aagattctag agctagcgaa ttcgaattta aatcggatcc gcggccgcaa ggatctgcga
     3301 tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     3361 gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag
     3421 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     3481 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct
     3541 tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg
     3601 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt
     3661 ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct
     3721 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt
     3781 tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga
     3841 cgccaccatg gagagcgacg agagcggcct gcccgccatg gagatcgagt gccgcatcac
     3901 cggcaccctg aacggcgtgg agttcgagct ggtgggcggc ggagagggca cccccaagca
     3961 gggccgcatg accaacaaga tgaagagcac caaaggcgcc ctgaccttca gcccctacct
     4021 gctgagccac gtgatgggct acggcttcta ccacttcggc acctacccca gcggctacga
     4081 gaaccccttc ctgcacgcca tcaacaacgg cggctacacc aacacccgca tcgagaagta
     4141 cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc tacgaggccg gccgcgtgat
     4201 cggcgacttc aaggtggtgg gcaccggctt ccccgaggac agcgtgatct tcaccgacaa
     4261 gatcatccgc agcaacgcca ccgtggagca cctgcacccc atgggcgata acgtgctggt
     4321 gggcagcttc gcccgcacct tcagcctgcg cgacggcggc tactacagct tcgtggtgga
     4381 cagccacatg cacttcaaga gcgccatcca ccccagcatc ctgcagaacg ggggccccat
     4441 gttcgccttc cgccgcgtgg aggagctgca cagcaacacc gagctgggca tcgtggagta
     4501 ccagcacgcc ttcaagaccc ccatcgcctt cgccagatcc cgcgctcagt cgtccaattc
     4561 tgccgtggac ggcaccgccg gacccggctc caccggatct cgcgagggca gaggaagtct
     4621 tctaacatgc ggtgacgtgg aggagaatcc cggccctatg accgagtaca agcccacggt
     4681 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     4741 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     4801 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     4861 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     4921 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     4981 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     5041 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     5101 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     5161 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     5221 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaaat caacctctgg attacaaaat
     5281 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5341 tgctttaatg cctttgtatc agttaacttg tttattgcag cttataatgg ttacaaataa
     5401 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5461 ttgtccaaac tcatcaatgt atcttatcat gtctggaatt gactcaaatg atgtcaatta
     5521 gtctatcaga agctatctgg tctcccttcc gggggacaag acatccctgt ttaatattta
     5581 aacagcagtg ttcccaaact gggttcttat atcccttgct ctggtcaacc aggttgcagg
     5641 gtttcctgtc ctcacaggaa cgaagtccct aaagaaacag tggcagccag gtttagcccc
     5701 ggaattgact ggattccttt tttagggccc attggtatgg ctttttcccc gtatcccccc
     5761 aggtgtctgc aggctcaaag agcagcgaga agcgttcaga ggaaagcgat cccgtgccac
     5821 cttccccgtg cccgggctgt ccccgcacgc tgccggctcg gggatgcggg gggagcgccg
     5881 gaccggagcg gagccccggg cggctcgctg ctgcccccta gcgggggagg gacgtaatta
     5941 catccctggg ggctttgggg gggggctgtc cctgatatct ataacaagaa aatatatata
     6001 taataagtta tcacgtaagt agaacatgaa ataacaatat aattatcgta tgagttaaat
     6061 cttaaaagtc acgtaaaaga taatcatgcg tcattttgac tcacgcggtc gttatagttc
     6121 aaaatcagtg acacttaccg cattgacaag cacgcctcac gggagctcca agcggcgact
     6181 gagatgtcct aaatgcacag cgacggattc gcgctattta gaaagagaga gcaatatttc
     6241 aagaatgcat gcgtcaattt tacgcagact atctttctag ggttaatcta gctgcatcag
     6301 gatcatatcg tcgggtcttt tttccggctc agtcatcgcc caagctggcg ctatctgggc
     6361 atcggggagg aagaagcccg tgccttttcc cgcgaggttg aagcggcatg gaaagagttt
     6421 gccgaggatg actgctgctg cattgacgtt gagcgaaaac gcacgtttac catgatgatt
     6481 cgggaaggtg tggccatgca cgcctttaac ggtgaactgt tcgttcaggc cacctgggat
     6541 accagttcgt cgcggctttt ccggacacag ttccggatgg tcagcccgaa gcgcatcagc
     6601 aacccgaaca ataccggcga cagccggaac tgccgtgccg gtgtgcagat taatgacagc
     6661 ggtgcggcgc tgggatatta cgtcagcgag gacgggtatc ctggctggat gccgcagaaa
     6721 tggacatgga taccccgtga gttacccggc gggcgcgctt ggcgtaatca tggtcatagc
     6781 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6841 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6901 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6961 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7021 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7081 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7141 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7201 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7261 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7321 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7381 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7441 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7501 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7561 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7621 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7681 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7741 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7801 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7861 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7921 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7981 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8041 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8101 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8161 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8221 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8281 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8341 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8401 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8461 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8521 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8581 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8641 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8701 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8761 gaataagggc gacacggaaa tgttgaatac tcat
//
